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Depletion of nuclear LINE1 RNA in mouse ESCs and embryos
LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote m...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8361322/ https://www.ncbi.nlm.nih.gov/pubmed/34409306 http://dx.doi.org/10.1016/j.xpro.2021.100726 |
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author | Percharde, Michelle Lin, Chih-Jen Ramalho-Santos, Miguel |
author_facet | Percharde, Michelle Lin, Chih-Jen Ramalho-Santos, Miguel |
author_sort | Percharde, Michelle |
collection | PubMed |
description | LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote microinjection of LINE antisense oligos, followed by RNA FISH validation. This protocol can be used in development, as well as other cell types where LINE1 is believed to be expressed. For complete information on the use and execution of this protocol, please refer to Percharde et al. (2018). |
format | Online Article Text |
id | pubmed-8361322 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-83613222021-08-17 Depletion of nuclear LINE1 RNA in mouse ESCs and embryos Percharde, Michelle Lin, Chih-Jen Ramalho-Santos, Miguel STAR Protoc Protocol LINE1 is the most active and abundant family of retrotransposons; it is implicated in a number of pathologies, as well as in early embryo development. We present a protocol to specifically knockdown LINE1 in mouse embryonic stem cells and embryos, including details for the nucleofection and zygote microinjection of LINE antisense oligos, followed by RNA FISH validation. This protocol can be used in development, as well as other cell types where LINE1 is believed to be expressed. For complete information on the use and execution of this protocol, please refer to Percharde et al. (2018). Elsevier 2021-08-09 /pmc/articles/PMC8361322/ /pubmed/34409306 http://dx.doi.org/10.1016/j.xpro.2021.100726 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Percharde, Michelle Lin, Chih-Jen Ramalho-Santos, Miguel Depletion of nuclear LINE1 RNA in mouse ESCs and embryos |
title | Depletion of nuclear LINE1 RNA in mouse ESCs and embryos |
title_full | Depletion of nuclear LINE1 RNA in mouse ESCs and embryos |
title_fullStr | Depletion of nuclear LINE1 RNA in mouse ESCs and embryos |
title_full_unstemmed | Depletion of nuclear LINE1 RNA in mouse ESCs and embryos |
title_short | Depletion of nuclear LINE1 RNA in mouse ESCs and embryos |
title_sort | depletion of nuclear line1 rna in mouse escs and embryos |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8361322/ https://www.ncbi.nlm.nih.gov/pubmed/34409306 http://dx.doi.org/10.1016/j.xpro.2021.100726 |
work_keys_str_mv | AT perchardemichelle depletionofnuclearline1rnainmouseescsandembryos AT linchihjen depletionofnuclearline1rnainmouseescsandembryos AT ramalhosantosmiguel depletionofnuclearline1rnainmouseescsandembryos |